Maternal Prkce expression in mature oocytes is critical for the first cleavage facilitating maternal-to-zygotic transition in mouse early embryos.
Maternal Prkce expression in mature oocytes is critical for the first cleavage facilitating maternal-to-zygotic transition in mouse early embryos.
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成熟卵母细胞中母体 Prkce 的表达对于促进小鼠早期胚胎中母体向合子转变的第一次卵裂至关重要
DOI：
10.1111/cpr.13231
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发表时间：
2022-06
影响因子：
8.5
通讯作者：
中科院分区：
文献类型：
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早期胚胎发育依赖于在母体到合子过渡期间储存在卵母细胞中的母体信息的调节。以往的研究报道了不同近交系小鼠卵母细胞能力的差异。本研究旨在通过比较来自高或低能力小鼠品系的卵母细胞的转录组来鉴定负责早期胚胎发育的母体转录物。 使用微阵列、RNA测序、真实的实时定量PCR（RT-qPCR）分析、蛋白质印迹和免疫荧光法对来自不同小鼠品系卵母细胞的体外受精胚胎进行分析。使用Prkce敲除小鼠分析一个候选基因Prkce，然后进行cRNA拯救实验。FVB/NJ（85.1%和82.0%）和DBA/2 J（79.6%和76.7%）近交系小鼠的受精率和2细胞率显著高于MRL/lpr（39.9%和35.8%）和129 S3（35.9%和36.6%）品系。记录了39个差异表达基因（DEG），其中9个通过RT-qPCR进一步验证。Prkce基因敲除小鼠的2细胞率降低（Prkce +/+ 80.1% vs. Prkce −/− 32.4%），可通过Prkce cRNA注射进行挽救（2细胞率达到76.7%）。整体转录分析显示，敲除小鼠中有143个DEG，主要由细胞周期调控基因组成。成熟卵母细胞中母体信息（如Prkce）的转录水平与选定近交系小鼠品系中不同的2细胞率相关。Prkce转录水平可以作为表征高质量成熟卵母细胞的潜在生物标志物。卵母细胞的能力是不同的小鼠品系之间的变量，最有可能是由于在MZT期间的卵母细胞中的母体转录本的差异丰度。一个候选基因Prkce在促进MZT中起着重要作用，特别是在第一次卵裂期间。Prkce转录本的表达水平似乎与不同小鼠品系中不同的卵母细胞能力相关。
Early embryo development is dependent on the regulation of maternal messages stored in the oocytes during the maternal‐to‐zygote transition. Previous studies reported variability of oocyte competence among different inbred mouse strains. The present study aimed to identify the maternal transcripts responsible for early embryonic development by comparing transcriptomes from oocytes of high‐ or low‐ competence mouse strains. In vitro fertilization embryos from oocytes of different mouse strains were subject to analysis using microarrays, RNA sequencing, real‐time quantitative PCR (RT‐qPCR) analysis, Western blotting, and immunofluorescence. One candidate gene, Prkce, was analysed using Prkce knockout mice, followed by a cRNA rescue experiment. The fertilization and 2‐cell rate were significantly higher for FVB/NJ (85.1% and 82.0%) and DBA/2J (79.6% and 76.7%) inbred mouse strains than those for the MRL/lpr (39.9% and 35.8%) and 129S3 (35.9% and 36.6%) strains. Thirty‐nine differentially expressed genes (DEGs) were noted, of which nine were further verified by RT‐qPCR. Prkce knockout mice showed a reduced 2‐cell rate (Prkce +/+ 80.1% vs. Prkce −/− 32.4%) that could be rescued by Prkce cRNA injection (2‐cell rate reached 76.7%). Global transcriptional analysis revealed 143 DEGs in the knockout mice, which were largely composed of genes functioning in cell cycle regulation. The transcription level of maternal messages such as Prkce in mature oocytes is associated with different 2‐cell rates in select inbred mouse strains. Prkce transcript levels could serve as a potential biomarker to characterize high‐quality mature oocytes. Oocyte competence is variable among different mouse strains, most likely due to the differential abundance of maternal transcripts in the oocyte during MZT. One candidate gene, Prkce, plays an essential role in facilitating MZT, especially during the first cleavage. The expression level of the Prkce transcript appears to be associated with different oocyte competence in different mice strains.
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